{"doi":"10.1101/2022.04.12.488038","title":"A streamlined tandem tip-based workflow for sensitive nanoscale phosphoproteomics","abstract":"Abstract Effective phosphoproteome of nanoscale sample analysis remains a daunting task, primarily due to significant sample loss associated with non-specific surface adsorption during enrichment of low stoichiometric phosphopeptide. We developed a novel tandem tip phosphoproteomics sample preparation method that is capable of sample cleanup and enrichment without additional sample transfer, and its integration with our recently developed SOP (Surfactant-assisted One-Pot sample preparation) and iBASIL (improved Boosting to Amplify Signal with Isobaric Labeling) approaches provides a streamlined workflow enabling sensitive, high-throughput nanoscale phosphoproteome measurements. This approach significantly reduces both sample loss and processing time, allowing the identification of >3,000 (>9,500) phosphopeptides from 1 (10) µg of cell lysate using the label-free method without a spectral library. It also enabled precise quantification of ∼600 phosphopeptides from 100 cells sorted by FACS (single-cell level input for the enriched phosphopeptides) and ∼700 phosphopeptides from human spleen tissue voxels with a spatial resolution of 200 µm (equivalent to ∼100 cells) in a high-throughput manner. The new workflow opens avenues for phosphoproteome profiling of mass-limited samples at the low nanogram level.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2022,"id":301120,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9492,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":291294,"name":"Yi-Ting Wang","orcid":"0000-0001-7509-2016","position":1,"is_corresponding":false},{"id":250402,"name":"Chuan‐Chih Hsu","orcid":"0000-0002-7100-1401","position":2,"is_corresponding":false},{"id":869704,"name":"Reta Birhanu Kitata","orcid":"0000-0002-8973-7679","position":3,"is_corresponding":false},{"id":232132,"name":"Rosalie Chu","orcid":"0000-0001-7428-7647","position":4,"is_corresponding":false},{"id":943213,"name":"Marija Veličković","orcid":"0000-0003-3664-5719","position":5,"is_corresponding":false},{"id":232133,"name":"Rui Zhao","orcid":"0000-0002-6602-6583","position":6,"is_corresponding":false},{"id":992292,"name":"Sarai M. Williams","orcid":null,"position":7,"is_corresponding":false},{"id":630561,"name":"William Chrisler","orcid":"0000-0002-3829-2454","position":8,"is_corresponding":false},{"id":992293,"name":"Marda L. Jorgensen","orcid":null,"position":9,"is_corresponding":false},{"id":60416,"name":"Ronald Moore","orcid":"0000-0003-2806-2855","position":10,"is_corresponding":false},{"id":4911,"name":"Ying Zhu","orcid":"0000-0002-5416-0566","position":11,"is_corresponding":false},{"id":60421,"name":"Karin Rodland","orcid":"0000-0001-7070-6541","position":12,"is_corresponding":false},{"id":45123,"name":"Richard Smith","orcid":"0000-0002-2381-2349","position":13,"is_corresponding":false},{"id":477240,"name":"Clive Wasserfall","orcid":"0000-0002-3522-8932","position":14,"is_corresponding":false},{"id":291295,"name":"Tujin Shi","orcid":"0000-0002-5592-3588","position":15,"is_corresponding":false},{"id":232135,"name":"Tao Liu","orcid":"0000-0001-9529-6550","position":16,"is_corresponding":false},{"id":281289,"name":"Chia‐Feng Tsai","orcid":"0000-0002-6514-6911","position":0,"is_corresponding":true}],"reference_count":65,"raw_metadata":null,"created_at":"2026-07-19T00:31:57.812980Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}